Método de análise computadorizado automatizado para quantificação celular em membranas de fibrina rica em plaquetas (PRF)

Abstract

The aim of this study was to compare different methods of histomorphometric analysis and to introduce a new automated method for cell quantification in Platelet Rich Fibrin (PRF) membranes. Methodology: 5 volunteers took part in the study, in which 3 venipunctures were performed at different times. In each case, 16mL of blood was collected and separated into 2 plastic tubes without additives. All the tubes were centrifuged using the L-PRF protocol (700g/12minutes) and, after centrifugation, the PRF was aspirated and dispensed into glass containers. The PRF clot obtained was pressed in PRF-BOX and the membranes generated were fixed in formaldehyde solution buffered with PBS. The membranes were then processed for paraffin embedding. The histological sections obtained were stained with hematoxylin and eosin (HE) and cell quantification was carried out using two approaches: 1) conventional histomorphometric analysis and 2) automated analysis, with the creation of a plug-in for ImageJ software. The time required to perform the analysis using each method was also measured. The results were submitted to the Bland-Altman statistical test and then to the linear regression test to check for proportion bias (the tendency for values to be above or below the mean). Results: no statistically significant difference was found in the number of cells obtained by the two methods (p = 0.1). The automated method reduced working time compared to the conventional method (p<0.01). Conclusion: The automated method developed was effective in histomorphometric analysis for cell quantification in PRF membranes and showed a significant reduction in working time.